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Fixable cytoplasmic stains for monitoring cell division by flow cytometry. The dyes can also be used to track cell populations in co-culture.
ViaFluor® SE Cell Proliferation Kits use amine-reactive dyes to covalently label cells throughout the cell cytoplasm and intracellular compartments for fixable fluorescent staining. Cell proliferation dyes are commonly used to monitor cell division by flow cytometry. The dyes also can be used to stably label cells to image cell morphology, or to track cell populations in mixed co-culture experiments.
ViaFluor® SE dyes are membrane-permeant compounds that are initially non-fluorescent esters, but are converted to fluorescent dyes by intracellular esterases and will covalently react with amine groups on intracellular proteins at the same time, forming fluorescent conjugates that are retained in the cell. Immediately after staining, a single bright fluorescent population will be detected by flow cytometry. With each cell division, daughter cells inherit roughly half of the fluorescent label, allowing the number of cell divisions that occur after labeling to be detected by the appearance of successively dimmer fluorescent peaks on a flow cytometry histogram compared to cells analyzed immediately after staining. Thus, cell proliferation dyes can be used to track multiple cell divisions of cells grown in culture or injected in vivo after labeling with the ViaFluor® SE dye.
The number of assays that can be performed per kit depends on the dye concentration used (see the product protocol for more information). When used at 1 uM to label 106 cells in one mL, each dye vial can be used for 90-100 labelings.

Human PBMCs were stained with ViaFluor® 405 (left), ViaFluor® 488 (center), or ViaFluor® 650 (right). Cells were stimulated with Dynabeads® Human T-Activator CD3/CD28 beads and 100 ng/mL IL-2. Cells were analyzed 4 days post-induction. CD3+ T-cells are shown. Unstimulated cells (dark peaks) and unstained cells (gray) are shown for comparison.
ViaFluor® SE dyes also can be used for imaging cell morphology or identifying cells in co-culture by microscopy. Because the staining is non-toxic and well-retained, it can be used for imaging live cells over time. See our Tech Tip: Using ViaFluor® SE Stains for Cell Tracing and Co-Culture.
All three ViaFluor® SE dyes can stain gram-positive bacteria, but not gram-negative bacteria. ViaFluor® CFSE stains the cytoplasm in yeast, but ViaFluor® 405 & ViaFluor® 488 stain the yeast cell periphery. See our Cellular Stains Table for more information on how our dyes stain various organisms.
Cell Division | Catalog No. | Ex/Em (nm) | Flow detection | Features |
|---|---|---|---|---|
| ViaFluor® 405 SE Cell Proliferation Kit | 30068 | 408/452 | Pacific Blue® | • Track cell division by dye dilution using flow cytometry • ViaFluor® 488 is a unique, improved green dye to replace CFSE • ViaFluor® 405 replaces CellTrace™ Violet • ViaFluor® 650 replaces CellTrace™ Far Red |
| ViaFluor® 488 SE Cell Proliferation Kit | 30086 | 493/532 | FITC | |
| ViaFluor® 650 SE Cell Proliferation Kit | 30139 | 653/682 | APC | |
| ViaFluor® CFSE Cell Proliferation Kit | 30050 | 495/519 | FITC |
It has been reported in publications that concentrations of serum above 10% in the assay may affect the results.
See the following publications for more information
Our ViaFluor® SE Cell Proliferation assay is a dye dilution assay for cell division, like CFSE and CellTrace™ Violet from Thermo. This type of assay is commonly used to measure lymphocyte proliferative responses in culture and in vivo (if the labeled cells are injected back into mice). It requires flow cytometry to analyze and allows you to count how many cell divisions have occurred in the labeled cells.
For more information and a typical procedure for using fluorescent ViaFluor® SE Dyes with PMBCs, which can easily be adapted for use with other cell types, please see our Tech Tip: Measuring Cell Division in PMBCs by Flow Cytometry
If flow cytometry is not an option, we offer absorbance-based and fluorescence-based microplate assays for quantitating cell numbers. These measure mitochondrial activity (resazurin/MTT/XTT) or intracellular esterase activity (calcein AM) as a readout of viable cell numbers. Please visit the Cell Viability and Apoptosis technology page for more information.
The ATP-Glo™ assay is a luminescence assay for cellular ATP levels, which are proportional to the number of live cells. This assay requires a luminometer.
CellTrace is a trademark of Thermo Fisher Scientific.
Our Resazurin Cell Viability Assay (Cat. No. 30025) has red fluorescence (Ex/Em 530-560/590 nm), and is specifically designed for microplate reader. It is an economical, easy-to-use, and homogeneous (no-wash) assay for quantifying live cells. It is similar to alamarBlue®, PrestoBlue®, and CellTiter-Blue®.
The Calcein AM Cell Viability Assay (Cat. No. 30026) has green fluorescence (Ex/Em 485/530 nm), and also works well for microplate reader. This assay requires culture medium to be removed from cells before adding the viability dye in buffer. We also offer the Viability/Cytotoxicity Assay for Animal Live & Dead Cells, which combines calcein-AM with the fluorescent dead cell stain EthD-III, and is compatible with microplate reader.